Recombinant
RabMAb

Anti-CD146 antibody [EPR3208] (ab75769)

Overview

  • Product name
    Anti-CD146 antibody [EPR3208]
    See all CD146 primary antibodies
  • Description
    Rabbit monoclonal [EPR3208] to CD146
  • Tested applications
    Suitable for: ICC/IF, IHC-Fr, WB, IHC-P, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human CD146 aa 600 to the C-terminus.
    Database link: P43121

  • Positive control
    • WB: A375, HUVEC and B16-F0 cell lysate; Human fetal artery lysate and Rat placenta ICC/IF: Murine bone marrow cell lysates. IHC-Fr: Mouse spleen tissue. IHC-P: Melanoma, breast carcinoma vessel, urinary bladder transitional carcinoma vessel, glioma vessel, normal tonsil and normal spleen tissue. FC: A375 and HUVEC cells
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab75769 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF 1/100 - 1/250.
IHC-Fr 1/250.
WB 1/1000. Predicted molecular weight: 72 kDa.Can be blocked with CD146 peptide (ab219161).

For unpurified use at 1/10000 - 1/50000.

IHC-P 1/250 - 1/500. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

Flow Cyt 1/20 - 1/80.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Target

  • Function
    Plays a role in cell adhesion, and in cohesion of the endothelial monolayer at intercellular junctions in vascular tissue. Its expression may allow melanoma cells to interact with cellular elements of the vascular system, thereby enhancing hematogeneous tumor spread. Could be an adhesion molecule active in neural crest cells during embryonic development. Acts as surface receptor that triggers tyrosine phosphorylation of FYN and PTK2, and a transient increase in the intracellular calcium concentration.
  • Tissue specificity
    Detected in endothelial cells in vascular tissue throughout the body. May appear at the surface of neural crest cells during their embryonic migration. Appears to be limited to vascular smooth muscle in normal adult tissues. Associated with tumor progression and the development of metastasis in human malignant melanoma. Expressed most strongly on metastatic lesions and advanced primary tumors and is only rarely detected in benign melanocytic nevi and thin primary melanomas with a low probability of metastasis.
  • Sequence similarities
    Contains 3 Ig-like C2-type (immunoglobulin-like) domains.
    Contains 2 Ig-like V-type (immunoglobulin-like) domains.
  • Cellular localization
    Membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • A32 antigen antibody
    • CD 146 antibody
    • CD146 antibody
    • CD146 antigen antibody
    • Cell surface glycoprotein MUC18 antibody
    • Cell surface glycoprotein P1H12 antibody
    • Gicerin antibody
    • Mcam antibody
    • Melanoma adhesion molecule antibody
    • Melanoma associated antigen A32 antibody
    • Melanoma associated antigen MUC18 antibody
    • Melanoma associated glycoprotein MUC18 antibody
    • Melanoma cell adhesion molecule antibody
    • Melanoma-associated antigen A32 antibody
    • Melanoma-associated antigen MUC18 antibody
    • MelCAM antibody
    • MUC 18 antibody
    • MUC18 antibody
    • MUC18_HUMAN antibody
    • S endo 1 antibody
    • S endo 1 endothelial associated antigen antibody
    • S-endo 1 endothelial-associated antigen antibody
    see all

Anti-CD146 antibody [EPR3208] images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human spleen tissue labelling CD146 with purified ab75769 at 1/250. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a goat anti-rabbit IgG H&L (HRP) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  • All lanes : Anti-CD146 antibody [EPR3208] (ab75769) at 1/10000 dilution (purified)

    Lane 1 : A375 cell lysate
    Lane 2 : Human fetal artery lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

    Predicted band size : 72 kDa

    Blocking buffer and concentration: 5% NFDM/TBST.
    Diluting buffer and concentration: 5% NFDM /TBST.

  • Immunocytochemistry/Immunofluorescence analysis of A375 (human malignant melanoma) cells labelling CD146 with purified ab75769 at 1/250. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

    Control 1: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).

    Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500).

  • Flow Cytometry analysis of A375 (human malignant melanoma) cells labeling CD146 with unpurified ab75769 at 1/20 dilution (10ug/mL) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor®488) at 1/2000 dilution was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) was used as the unlabeled control.

  • Anti-CD146 antibody [EPR3208] (ab75769) at 1/10000 dilution (purified) + HUVEC cell lysate at 20 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

    Predicted band size : 72 kDa

    Blocking buffer and concentration: 5% NFDM/TBST.
    Diluting buffer and concentration: 5% NFDM /TBST.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of melanoma tissue labelling CD146 with unpurified ab75769 at 1/250. A HRP/AP polymerized secondary antibody was used.

  • Immunohistochemistry (Frozen sections) analysis of mouse spleen tissue labelling CD146 with unpurified ab75769 at 1/250. Tissue samples were fixed with acetone and blocked with 5% serum for 2 hours at 25°C. The sample was incubated with primary antibody at 4°C for 12 hours. An Alexa Fluor®488-conjugated Goat polyclonal to rabbit IgG (1/250) was used as secondary antibody. Nuclear staining was with DAPI (blue).

    See Abreview

  • Flow Cytometry analysis of HUVEC cells labelling CD146 with purified ab75769 at 1/50 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

  • Anti-CD146 antibody [EPR3208] (ab75769) at 1/10000 dilution (purified) + B16-F0 cell lysate at 20 µg/ml

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

    Predicted band size : 72 kDa

    Blocking buffer and concentration: 5% NFDM/TBST.
    Diluting buffer and concentration: 5% NFDM /TBST.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of breast carcinoma vessels tissue labelling CD146 with unpurified ab75769.

  • Anti-CD146 antibody [EPR3208] (ab75769) at 1/10000 dilution (purified) + Rat placenta lysate at 20 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

    Predicted band size : 72 kDa

    Blocking buffer and concentration: 5% NFDM/TBST.
    Diluting buffer and concentration: 5% NFDM /TBST.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of urinary bladder transitional carcinoma vessels tissue labelling CD146 with unpurified ab75769.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of glioma vessels tissue labelling CD146 with unpurified ab75769.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of normal tonsil tissue labelling CD146 with unpurified ab75769.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of normal spleen tissue labelling CD146 with unpurified ab75769.

References for Anti-CD146 antibody [EPR3208] (ab75769)

This product has been referenced in:
  • Liu Y  et al. Nrf2 Inhibits Periodontal Ligament Stem Cell Apoptosis under Excessive Oxidative Stress. Int J Mol Sci 18:N/A (2017). Read more (PubMed: 28513573) »
  • Jacques C  et al. Targeting the epigenetic readers in Ewing sarcoma inhibits the oncogenic transcription factor EWS/Fli1. Oncotarget 7:24125-40 (2016). IHC . Read more (PubMed: 27006472) »

See all 36 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Human brain pericytes)
Permeabilization
Yes - Triton X-100
Specification
Human brain pericytes
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 37°C
Fixative
Paraformaldehyde
Username

Radhika Menon

Verified customer

Submitted Jul 06 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Brain)
Permeabilization
No
Specification
Brain
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Sep 28 2016

Application
Immunohistochemistry (Frozen sections)
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Sample
Mouse Tissue sections (Dental pulp)
Specification
Dental pulp
Permeabilization
No
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Feb 12 2015

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 20°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate
Sample
Human Tissue sections (blood vessels)
Specification
blood vessels
Permeabilization
No
Fixative
Formaldehyde
Username

Mr. Florian Guilbaud

Verified customer

Submitted Jun 25 2014

Application
Western blot
Loading amount
37.5 µg
Gel Running Conditions
Reduced Denaturing (7% tris acetate gel)
Sample
Human Cell lysate - whole cell (Renal Cancer Cell Line 786-0)
Specification
Renal Cancer Cell Line 786-0
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Dr. Victoria Thompson

Verified customer

Submitted May 13 2014

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Antigen retrieval step
None
Sample
Human Tissue sections (tooth)
Specification
tooth
Permeabilization
No
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

Submitted Apr 04 2014

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Antigen retrieval step
None
Sample
Human Tissue sections (UBC)
Specification
UBC
Permeabilization
No
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Mar 26 2014

Application
Immunohistochemistry (Frozen sections)
Blocking step
NOVOCASTRA protein block as blocking agent as blocking agent as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 25°C
Sample
Mouse Tissue sections (E10.5 mouse embryo)
Specification
E10.5 mouse embryo
Permeabilization
Yes - PBS / 0.5% v/v Triton X-100 (Sigma)
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Oct 31 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (stromal)
Specification
stromal
Fixative
Paraformaldehyde
Permeabilization
Yes - Triton
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted May 03 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (colon)
Specification
colon
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Leica Bond Epitope Retrieval 2
Permeabilization
No
Username

Abcam user community

Verified customer

Submitted Mar 26 2013

1-10 of 36 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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